CN112540162A - Water quality biotoxicity detection method - Google Patents
Water quality biotoxicity detection method Download PDFInfo
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- CN112540162A CN112540162A CN202011339872.2A CN202011339872A CN112540162A CN 112540162 A CN112540162 A CN 112540162A CN 202011339872 A CN202011339872 A CN 202011339872A CN 112540162 A CN112540162 A CN 112540162A
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- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/18—Water
- G01N33/186—Water using one or more living organisms, e.g. a fish
- G01N33/1866—Water using one or more living organisms, e.g. a fish using microorganisms
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- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
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- Y02A20/20—Controlling water pollution; Waste water treatment
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Abstract
The invention discloses a water quality biotoxicity detection method, which comprises the following steps: collecting water to be detected, and performing dechlorination treatment on the water to be detected; preparing a microbial liquid; and (3) detecting biological toxicity: and detecting the initial luminescence amount of the microbial liquid, then putting the microbial liquid into the water quality to be detected, detecting the luminescence amount of the microbial liquid again, and calculating the light inhibition rate to realize the detection of the biotoxicity of the water quality to be detected. The invention has the beneficial effects that: the water quality biotoxicity detection method can present the water quality problem in a short time, improves timeliness, sensitivity and accuracy of biological monitoring and early warning, is less in time consumption, simple to operate and accurate in result, provides support for sudden water quality pollution and a water treatment technology of a water plant, reduces risks caused by drinking water exposure, and guarantees water supply quality safety.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a water quality biotoxicity detection method.
Background
With the development of modern industry, the use of chemical substances is increasing day by day, so that aquatic ecosystems on which human beings live are polluted more and more seriously, and sudden environmental pollution accidents occur, such as sudden water quality change caused by artificial toxicity input and natural disasters, especially the pollution to environmental water bodies caused by accidents occurring in the production, storage and transportation processes of petrochemical materials, products and toxic and harmful dangerous goods. This requires that we deal with various sudden environmental pollution accidents quickly and minimize various economic losses or social influences. For decades, the sensitivity of various physicochemical analytical means has been increasing, and most researchers have been concerned about the toxic effect of single pollutants on organisms and ecosystems, but organisms in the environment are often exposed to mixed systems in which multi-component pollutants coexist, rather than simple single systems. The toxic effect produced by the mixture system is the combined result of the antagonistic, additive, synergistic or inhibitory effects of all the component pollutants, and even when the individual components in the mixture system are at non-toxic effect concentrations, the components still contribute to the overall toxic effect of the mixture. Therefore, it is very urgent and necessary to develop a new effective method for rapidly and accurately evaluating the toxicity of various pollutants.
The luminescent bacteria toxicity test is one of the classical comprehensive biological toxicity evaluation methods, and because of the characteristics of short growth period of bacteria, sensitivity to environmental changes and the like, the change degree of the luminescent intensity is theoretically linearly related to the mass concentration of the toxicant. Bacterial tests represented by luminescent bacteria are widely applied at home and abroad, and the luminescent bacteria method is taken as a standard toxicity test method in many countries. At present, a luminous bacteria toxicity instrument is also applied to water quality early warning detection and is used as one of powerful barriers for water quality safety.
Disclosure of Invention
The invention aims to provide a water quality biotoxicity detection method which can improve water quality biotoxicity detection and can present water quality conditions in a short time.
In order to achieve the purpose, the invention adopts the following technical scheme:
a water quality biotoxicity detection method comprises the following steps:
(1) collecting water to be detected, and performing dechlorination treatment on the water to be detected;
(2) preparing a microbial liquid;
(3) and (3) detecting biological toxicity: and detecting the initial luminescence amount of the microbial liquid, then putting the microbial liquid into the water quality to be detected, detecting the luminescence amount of the microbial liquid again, and calculating the light inhibition rate to realize the detection of the biotoxicity of the water quality to be detected.
As a preferred embodiment, in the step (1), the dechlorination treatment is to treat the water to be detected through a dechlorination filter, preferably, the dechlorination filter is filled with a dechlorination agent, a liquid inlet is formed at the top end of the dechlorination filter, and a liquid outlet is formed at the bottom of the dechlorination filter.
In the above method for detecting biotoxicity of water, as a preferred embodiment, the dechlorinating agent is bismuth oxide with purity of more than 99.5%.
The bismuth oxide is generally bismuth trioxide, can remove more than 80% of chlorine in water, and can be recycled by alkali washing.
As a preferred embodiment, the method for detecting the biotoxicity of the water quality comprises the following steps: inoculating luminous bacteria into culture medium, and dispersing thallus with the dispersion liquid to obtain microbial liquid when the luminous bacteria is cultured to logarithmic phase.
In the above method for detecting water biotoxicity, as a preferred embodiment, the luminescent bacteria are vibrio fischeri or/and vibrio qinghaiensis.
The vibrio fischeri is a bacterium capable of emitting visible fluorescence, the normal luminous bacterium can emit blue-green visible light with the wavelength of 450-.
The vibrio qinghaiensis is a freshwater luminescent bacterium, can continuously and stably emit blue-green light (with the maximum emission wavelength of 485nm), can be quickly inhibited to emit light once encountering toxic and harmful substances, and has a corresponding relation between the light emission inhibition degree and the toxicity and concentration of the toxic and harmful substances.
As a preferred embodiment, the culture medium is LB solid culture medium containing 20-25ug/L kanamycin.
In the above-mentioned method for detecting the biotoxicity in water, as a preferred embodiment, the culture medium is LB solid medium containing 22ug/L kanamycin.
The water quality biotoxicity detection method is a preferable embodiment, the luminous bacteria are inoculated on the culture medium, the culture is carried out for 15-20h at the temperature of 20-25 ℃, and after the culture is finished, the lawn is flushed from the culture medium by using normal saline to obtain the microbial liquid.
In the above method for detecting water biotoxicity, as a preferred embodiment, in step (3), the microbial liquid is put into the water to be detected, and the luminescence amount of the microbial liquid is detected again after 15-30 min.
In the above-mentioned method for detecting biotoxicity in water, as a preferred embodiment, in the step (3), the light-suppressing rate is calculated by I ═ (R0-R)/R0 × 100%, where R0 is the initial luminescence amount of the microbial liquid, R is the luminescence amount of the microbial liquid after being put into water, and I is the light-suppressing rate.
Compared with the prior art, the invention has the beneficial effects that: the water quality biotoxicity detection method can present the water quality problem in a short time, improves timeliness, sensitivity and accuracy of biological monitoring and early warning, is less in time consumption, simple to operate and accurate in result, provides support for sudden water quality pollution and a water treatment technology of a water plant, reduces risks caused by drinking water exposure, and guarantees water supply quality safety.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be described in detail below. It is to be understood that the described embodiments are merely a few embodiments of the invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the examples given herein without any inventive step, are within the scope of the present invention.
Example 1
A water quality biotoxicity detection method comprises the following steps:
(1) collecting water to be detected, and performing dechlorination treatment on the water to be detected through a dechlorination filter;
as a preferred embodiment, the chlorine removal filter is filled with a chlorine removal agent, the top end of the chlorine removal filter is provided with a liquid inlet, and the bottom of the chlorine removal filter is provided with a liquid outlet; the water to be detected flows in from the liquid inlet of the dechlorination filter and flows out from the liquid outlet, so that the dechlorination of the water to be detected is realized;
as a preferred embodiment, the dechlorination agent is bismuth oxide with the purity of more than 99.5 percent, and the bismuth oxide can remove more than 80 percent of chlorine in water, thereby eliminating the influence of the chlorine on the biotoxicity detection of the water quality and ensuring that the detection result is more accurate;
(2) preparing a microbial liquid:
inoculating luminescent bacteria Vibrio qinghaiensis into LB solid culture medium containing 20ug/L kanamycin, culturing at 20 deg.C for 20 hr, and flushing thallus Porphyrae from the culture medium with normal saline when the luminescent bacteria culture reaches logarithmic phase to obtain microorganism solution;
(3) and (3) detecting biological toxicity: the initial luminescence of the detected microbial solution is recorded as R0Then, the microbial liquid is put into the water quality to be detected, the light-emitting quantity of the microbial liquid is detected again after 15min and is recorded as R, and the light inhibition rate I is calculated according to the method that I is recorded as I ═(R0-R)/R0X 100 percent, and realizes the detection of the biotoxicity of the water quality to be detected.
Example 2
A water quality biotoxicity detection method comprises the following steps:
(1) collecting water to be detected, and performing dechlorination treatment on the water to be detected through a dechlorination filter;
as a preferred embodiment, the chlorine removal filter is filled with a chlorine removal agent, the top end of the chlorine removal filter is provided with a liquid inlet, and the bottom of the chlorine removal filter is provided with a liquid outlet; the water to be detected flows in from the liquid inlet of the dechlorination filter and flows out from the liquid outlet, so that the dechlorination of the water to be detected is realized;
as a preferred embodiment, the dechlorination agent is bismuth oxide with the purity of more than 99.5 percent, and the bismuth oxide can remove more than 80 percent of chlorine in water, thereby eliminating the influence of the chlorine on the biotoxicity detection of the water quality and ensuring that the detection result is more accurate;
(2) preparing a microbial liquid:
inoculating luminescent bacteria of Vibrio fischeri or Vibrio harveyi and Vibrio qinghaiensis into LB solid culture medium containing 22ug/L kanamycin, culturing at 22 deg.C for 18h, and flushing thallus Porphyrae from the culture medium with normal saline when the luminescent bacteria is cultured to logarithmic phase to obtain microbial solution;
(3) and (3) detecting biological toxicity: the initial luminescence of the detected microbial solution is recorded as R0Then, the microbial liquid is put into the water quality to be detected, the light-emitting quantity of the microbial liquid is detected again after 25min and is recorded as R, and the light inhibition rate I is calculated by the method that I is (R)0-R)/R0X 100 percent, and realizes the detection of the biotoxicity of the water quality to be detected.
Example 3
A water quality biotoxicity detection method comprises the following steps:
(1) collecting water to be detected, and performing dechlorination treatment on the water to be detected through a dechlorination filter;
as a preferred embodiment, the chlorine removal filter is filled with a chlorine removal agent, the top end of the chlorine removal filter is provided with a liquid inlet, and the bottom of the chlorine removal filter is provided with a liquid outlet; the water to be detected flows in from the liquid inlet of the dechlorination filter and flows out from the liquid outlet, so that the dechlorination of the water to be detected is realized;
as a preferred embodiment, the dechlorination agent is bismuth oxide with the purity of more than 99.5 percent, and the bismuth oxide can remove more than 80 percent of chlorine in water, thereby eliminating the influence of the chlorine on the biotoxicity detection of the water quality and ensuring that the detection result is more accurate;
(2) preparing a microbial liquid:
inoculating luminescent bacteria, namely vibrio freundii, into an LB solid culture medium containing 25ug/L kanamycin, culturing at 20 ℃ for 20h, and flushing lawn from the culture medium by using normal saline when the luminescent bacteria are cultured to a logarithmic phase to prepare a microbial solution;
(3) and (3) detecting biological toxicity: the initial luminescence of the detected microbial solution is recorded as R0Then, the microbial liquid is put into the water quality to be detected, the light-emitting quantity of the microbial liquid is detected again after 30min and is recorded as R, and the light inhibition rate I is calculated by the method that I is (R)0-R)/R0X 100 percent, and realizes the detection of the biotoxicity of the water quality to be detected.
The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims (10)
1. A water quality biotoxicity detection method is characterized by comprising the following steps:
(1) collecting water to be detected, and performing dechlorination treatment on the water to be detected;
(2) preparing a microbial liquid;
(3) and (3) detecting biological toxicity: and detecting the initial luminescence amount of the microbial liquid, then putting the microbial liquid into the water quality to be detected, detecting the luminescence amount of the microbial liquid again, and calculating the light inhibition rate to realize the detection of the biotoxicity of the water quality to be detected.
2. The method for detecting the biotoxicity of the water quality as claimed in claim 1, wherein in the step (1), the dechlorination treatment is to treat the water to be detected through a dechlorination filter, preferably, the dechlorination filter is filled with a dechlorination agent, a liquid inlet is arranged at the top end of the dechlorination filter, and a liquid outlet is arranged at the bottom of the dechlorination filter.
3. The method as claimed in claim 2, wherein the dechlorinating agent is bismuth oxide with purity > 99.5%.
4. The method for detecting the biotoxicity of the water quality as claimed in claim 1, wherein the preparation method of the microbial liquid is as follows: inoculating luminous bacteria into culture medium, and dispersing thallus with the dispersion liquid to obtain microbial liquid when the luminous bacteria is cultured to logarithmic phase.
5. The method as claimed in claim 4, wherein the luminescent bacteria is Vibrio fischeri or/and Vibrio qinghaiensis.
6. A method for detecting biotoxicity in water as claimed in claim 4, wherein the culture medium is LB solid medium containing 20-25ug/L kanamycin.
7. A method as claimed in claim 4 wherein said culture medium is LB solid medium containing 22ug/L kanamycin.
8. A method as claimed in claim 4, wherein the luminous bacteria is inoculated onto the culture medium, cultured at 20-25 deg.C for 15-20h, and after the culture is finished, the lawn is flushed from the culture medium with normal saline to obtain the microbial solution.
9. A method for detecting the biotoxicity of water quality as claimed in claim 1, wherein in the step (3), the microbial liquid is put into the water quality to be detected, and the luminescence quantity of the microbial liquid is detected again after 15-30 min.
10. A method as claimed in claim 1, wherein in step (3), the light-inhibiting rate is calculated by the method of I ═ R (R)0-R)/R0X 100% where R0The initial luminescence amount of the microbial liquid, R is the luminescence amount of the microbial liquid after being put into water, and I is the light inhibition rate.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202011339872.2A CN112540162A (en) | 2020-11-25 | 2020-11-25 | Water quality biotoxicity detection method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114051957A (en) * | 2021-11-15 | 2022-02-18 | 江苏雅信昆成检测科技有限公司 | Large-scale daphnia of continuous-flow type test device |
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| CN114051957A (en) * | 2021-11-15 | 2022-02-18 | 江苏雅信昆成检测科技有限公司 | Large-scale daphnia of continuous-flow type test device |
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